Method for refining protamine sulfate

A technology of protamine sulfate and protamine, applied in the field of biomedicine, can solve the problem of high fatality rate

Inactive Publication Date: 2015-04-22
QINGDAO KANGYUAN PHARMA
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  • Abstract
  • Description
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Problems solved by technology

However, the literature and observations show that the incidence of protamine toxic reactions is high, the clinical manifestations are diverse, and the fatality ra

Method used

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Examples

Experimental program
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Embodiment 1

[0020] The protamine essence of salmon is crushed, extracted with water, filtered, and the filtrate is precipitated with ethanol to obtain the crude product of protamine sulfate, which is then subjected to the following operations:

[0021] (1) Protamine enzymatic hydrolysis process

[0022] Accurately weigh 50mg of protamine sulfate crude product, dissolve in 50ml CaCl 2 In the buffer solution, continue to stir and dissolve for about 1 hour until completely dissolved. Accurately weigh 5 mg of thermolysin, and add it into a fully dissolved protamine buffer to obtain a reaction solution with a protamine:thermolysin (w / w) ratio of 100:1. Filter and set aside.

[0023] (2) Separation and purification process

[0024] Separate and weigh Tris buffer salt and NaCl to prepare mobile phase A; 50mM Tris solution and mobile phase B:, 50mM Tris / 2MNaCl solution. The pH was adjusted to 7.5 with HCl. Install the heparin affinity chromatographic column on the fast protein chromatography...

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Abstract

The invention discloses a method for refining protamine sulfate. According to the technical scheme adopted by the invention, the method comprises the following steps: performing enzymolysis on the protamine sulfate, performing quick protein liquid-phase separation to purify enzymatic hydrolysate, desalting a protamine high-salt solution to obtain high-purity protamine sulfate. A polypeptide mixture with average molecular weight of 1100Da, obtained by performing enzymolysis on the protamine with thermolysin has the capacity of neutralizing heparin; immune response caused after injection is remarkably lowered, and the toxic and side effects are relieved.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for refining protamine sulfate by using suitable crude protamine as a raw material. Background technique [0002] Protamine (Latin name Protamine) is a basic protein that mainly exists as nuclear protamine bound to DNA in the mature sperm nucleus of fish (such as frogfish, squid, herring, etc.). [0003] Protamine was discovered in 1870, and it was not until 1940-1960 that research on its use as an antibacterial agent became prevalent. Protamine shows strong antibacterial ability in neutral and alkaline media, and has high thermal stability. It is still active when heated at 210°C for 1 hour. At the same time, the antibacterial range and food antiseptic range are both Wider, it has a strong inhibitory effect on Bacillus subtilis, Bacillus bacillus, Lactobacillus casei, Lactobacillus plantarum, lactic acid bacteria, mold, spore heat-resistant bacteria and Gram-positive bacteri...

Claims

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Application Information

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IPC IPC(8): C07K14/46C12P21/06C07K1/36C07K1/34C07K1/22
CPCC07K14/461C12P21/06
Inventor 刘乃山邓亚楠刘翠珍
Owner QINGDAO KANGYUAN PHARMA
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